Lucas-Hahn A, Niemann H
Institut für Tierzucht und Tierverhalten (FAL), Mariensee 3057 Neustadt 1, Germany.
Theriogenology. 1991 Oct;36(4):619-27. doi: 10.1016/0093-691x(91)90399-x.
Three experiments were conducted to investigate the effects of type of culture medium in freshly bisected bovine embryos and the effects of agar embedding and of 1.2 propanediol (PROH) as the cryoprotectant in frozen/thawed bisected bovine embryos on development in vitro. A total of 265 bovine embryos were used as controls or were microsurgically bisected and were cultured in vitro for 48 hours and development was determined 24 and 48 hours after the onset of culture. Whitten's medium supported more (P<0.05) intact and demi-embryos to grow to expanded blastocysts (92.9 and 73.1%, respectively) compared with Ham's F10 (43.8 and 26.3%, respectively) and PBS (53.8 and 12.5%, respectively). Embedding in agar and culture in Whitten's medium resulted in a higher (P<0.05) percentage of in vitro development of frozen/thawed demi-embryos after 24 hours than the freezing of nonembedded demi-embryos (44.1 versus 19.6%, respectively). This difference disappeared, however, after a 48 hours culture period (17.6 versus 11.8%, respectively). Following freezing in PROH, survival rates of 40 and 28%, respectively after 24 hours of culture were obtained for intact and demi-embryos. The respective percentages after 48 hours were 8.6 and 16%. Since neither embedding in agar nor the use of PROH as the cryoprotectant resulted in high survival rates of frozen/thawed demi-embryos in vitro, new freezing procedures are needed to overcome the sensitivity of demi-embryos to freezing and thawing.
进行了三项实验，以研究新鲜分割牛胚胎时培养基类型的影响，以及冷冻/解冻分割牛胚胎时琼脂包埋和1,2 - 丙二醇（PROH）作为冷冻保护剂对体外发育的影响。总共265个牛胚胎用作对照或经显微手术分割，体外培养48小时，并在培养开始后24小时和48小时确定发育情况。与Ham's F10（分别为43.8%和26.3%）和PBS（分别为53.8%和12.5%）相比，Whitten's培养基支持更多（P<0.05）的完整胚胎和半胚胎发育至扩张囊胚（分别为92.9%和73.1%）。琼脂包埋并在Whitten's培养基中培养，导致冷冻/解冻半胚胎在24小时后的体外发育百分比高于未包埋半胚胎的冷冻情况（分别为44.1%和19.6%）。然而，在48小时培养期后，这种差异消失了（分别为17.6%和11.8%）。在PROH中冷冻后，完整胚胎和半胚胎在培养24小时后的存活率分别为40%和28%。48小时后的相应百分比分别为8.6%和16%。由于琼脂包埋和使用PROH作为冷冻保护剂均未导致冷冻/解冻半胚胎在体外的高存活率，因此需要新的冷冻程序来克服半胚胎对冷冻和解冻的敏感性。
